DAGRS: Directed Antigonists to Cancer Cell Growth Signals

ABSTRACT

The present invention describes a unique method of treating cancer with the administration of an improved DAGRS™ construct which functions as a humanized agent specifically targeting cancer cells in vivo. A specific DAGRS™ is described constructed of a humanized drug delivery biologic, carboxyl to an Apoptin fragment consisting of Apoptin&#39;s proline-rich SH3-binding fragment, a spacer, and a MAP kinase (MAPK) phosphorylation site, in replacement of the SH3-binding domain at HIV-1 TAT&#39;s amino terminus. Apoptin is a viral protein with incumbent immunogenicity and toxicity in humans. Improved DAGRS™ constructs are described that replace the viral VP3 peptide with human AKT peptide or derivative, all equivalently spaced 11 amino acids from the initial proline to the beginning of the MAPK phosphorylation site, through which technology the DAGRS™ is fully humanized. DAGRS™ provide for improved bioavailability, enhanced specific activity, and low toxicity for in vivo treatment of cancer. DAGRS™ are a superior method for targeting any oncogene with an inhibitory peptide. 
     An algorithm for “humanization” is described through which human functional equivalent(s) to viral product(s) are identified by alignment of peptides anchored at each end by matching functional motifs that are spaced equivalently distant in the two aligned peptides. The algorithm totally disregards the primary amino acid composition of the spacer, and as such separates from current computer algorithms that prioritize primary amino acid alignments. Accounting for spacing dictates that functional domains be oriented correctly in three dimensions. The invention taught here can be developed into computer algorithms for rapidly identifying these anchored alignments, and thereafter developing safe humanized drugs from disruptive viral activities. Computers once taught the basic rules for anchoring equivalents, can improve on the basic algorithm through artificial intelligence to expand drug development.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/362,254, filed Jul. 14, 2016 and incorporated by reference in itsentirety.

FIELD OF THE INVENTION

The present invention relates to the field of oncogene-targetedtherapeutics in the treatment of cancer.

BACKGROUND

Cancer is among the leading causes of morbidity and mortality worldwidewith approximately 14 million new cases and 8.2 million cancer-relateddeaths in 2012 (WHO, World Cancer Report. Bernard W. Stewart andChristopher P. Wild, eds. 2014). The number of new cases is expected torise by about 75% over the next 2 decades coincident with an agingpopulation. One defining feature of cancer is the rapid creation ofabnormal cells that grow beyond their usual boundaries, and which canthen invade adjoining parts of the body and spread to other organs.Oncogenesis is the result of the interaction between genetic factors andexternal agents such as, but not limited to, ultraviolet radiation,asbestos tobacco smoke, or viral infection. Cancer-causing viralinfections such as HBV/HCV and HPV are responsible for up to 20% ofcancer deaths in low- and middle-income countries The transformationfrom normal cells into tumor cells is a multistage process, typically aprogression from a pre-cancerous lesion seeded by cancer stem cells, tomalignant tumors that metastasize to distant sites. Metastasis is theprimary cause of death for human cancers, while certain cancers thatrarely metastasize (basal cell carcinoma) are almost never fatal.

Current cancer treatments are dominated by invasive surgery, radiationtherapy and chemotherapy protocols, which are frequently ineffective andcan have potentially severe side-effects, non-specific toxicity and/orcause traumatizing changes to an individual's body image and/or qualityof life. One of the causes for the inadequacy of current cancertreatments is their lack of selectivity for affected tissues and cells.More selective cancer treatments would leave normal cells unharmed thusimproving outcome, side-effect profile and quality of life.

While significant advancements have been made, treatment of cancers bychemotherapy frequently results in severe side effects because thetherapy used is not specific to the cancer, killing non-cancerous cellsincluding hematopoietic cells critical to immune surveillance. Inaddition to standard chemotherapy and hormone replacement therapy, newclasses of therapies have emerged with directed oncolytic mechanisms.One approach targets either toxins or radioactive isotopes directly intothe cancers by coupling the oncolytic agent to monoclonal antibodies(MAb) directed against cancer antigen. Genentech's Kadcyla® is anexample of this kind of “smart-bomb” approved for the treatment ofbreast cancer. Another class are drugs like Gleevec® (Novartis) thatantagonize growth pathways specific to cancer cells, such as the bcr-abloncogene of chronic myelogenous leukemia targeted by Gleevec®. This isthe class of agent described in this invention, with the difference thatthis invention describes a biologic that is a drug delivery tool with aprogrammable cassette such that it can be theoretically targeted againstany oncogene. Other approaches are being designed directed againstgrowth pathways specific to cancer stem cells, which are the seeds forcancer metastasis to distant sites. This stem cell strategy is apreferred realization of this invention because it has been theorizedthat mutational escape of cancer stem cells is rare compared to cancertumor cells.

Other realizations of targeted cancer therapies are oncolytic viruses, atechnology based on the observation by Coley of spontaneous remissionsin certain blood cancers during severe systemic viral infections.Oncolytic viruses are currently approved for the treatment of certainblood dyscrasias and recurrent melanoma Kyprolis® (Amgen, Inc.) orcarfilzomib for injection for multiple myeloma, see U.S. Pat. No.9,315,542 and U.S. Pat. No. 9,309,283. Most recently an oncolyticpoliovirus developed at Duke Medical Center gained fast track approvalfor the treatment of recurrent glioblastoma. The Chicken Anemia Virus(CAV) has been noted to mediate oncolysis through its VP3 (“Apoptin”)protein, an observation that has remained in pre-clinical developmentowing to bioavailability and delivery issues.

At the present time, patients with recurrent cancer have few options oftreatment that offer extended quality of life. The regimented approachto cancer therapy has produced overall improvements in global survivaland morbidity rates. However, to the particular individual, theseimproved statistics do not necessarily correlate with an improvement intheir personal situation, or even to prolonged survival. When cancerrecurs after these consolidation therapies, it is almost always rapidlyfatal even when treated by any of the newer targeted agents.

An improved approach to treatment would be to design agents targeted toinhibiting oncogenes using Directed Antagonists to cancer Growth Signals(DAGRS™), with low toxicity and good bioavailability. This inventioncaptures the cancer-killing activity of certain oncolytic viruses in asimple peptide, sparing the toxicities associated with a multitude ofother viral proteins that are superfluous to oncolysis but a source oftoxicity. In the example of oncolyitc poliovirus for glioblastoma, theinvestigators had the profound head start to safety of an attenuatedpoliovirus that has been used safely for 60 years as a vaccine. Whileits approval for use in adult glioblastoma is a major advance,glioblastoma is also a disease of children. The coupling of thehistorically higher incidence of paralytic polio in children with theglobal immune suppression associated with cancers raises the concernthat the safety profile of the oncolytic poliovirus may not be nearly asgood in children as it is in adults.

The treatment of diverse cancers with the power and adaptability ofDAGRS™ is a major beneficial outcome that can derive from thisinvention. As one example, individual profiling of cancers is becomingcommonplace as a strategy to better tailor therapeutics to the uniquegenetics of the patient. Because in principle DAGRS™ can be targetedagainst any oncogene, a pair or even a trio of synergistic DAGRS™ couldbe administered to the cancer patient that precisely antagonize thatindividual's oncogene profile. As a second example, mutations have beendiscovered that render some fraction of cancers particularly susceptibleto specific directed oncolytics. Invariably the cancers under treatmentundergo mutational escape so that, while there is a short term benefitin tumor regression, the long term benefit in survival is mostfrequently marginal. Because the downstream escape pathways are limitedand reproducible, a DAGRS™ that targets and blocks the most commondownstream escape mechanism(s) could be administered along with anotherdirected oncolytic, thereby potentiating the efficacy of both.

SUMMARY

The present invention describes an improved composition and methods forthe treatment of cancer that incorporate the administration of asynthetic, genetically engineered Directed Antagonists to cancer GrowthSignals (DAGRS™) targeted at inhibiting an oncogene. Because DAGRS™ areconstructed from diverse small stretches of genetic material that aretiled together in a unique arrangement, DAGRS™ compositions of matterhave less than 50% homology to any naturally occurring biologic. DAGRshave the ability to deliver a biologic from outside the cell through thecytoplasm to the nucleus, and are engineered to bind to specific targetsthrough introduction of specific peptide fragments into a cassette thatlocks the peptide into a high affinity configuration. DAGRS™ can inprinciple be targeted against any oncogene. Preferred oncogene targetsillustrated in this invention are E2F and AKT, which are effectiveagainst many cancers in vitro. AKT is a particularly attractive targetbecause it is found to be mutated in approximately 80% of human cancers,its inhibition mediates a p53 independent apoptosis, and its specificactivity in G2 phase dividing (cancer) cells supports a good safetyprofile for normal cells. The CAV VP3 protein (“Apoptin”) mediatesoncolysis at least in part through AKT, and Apoptin has been shown tokill a wide variety of cancer cells but not normal cells in vitro.Apoptin has been coupled to Tat monomer as an in vitro delivery toolwith positive results. However, the toxicities inherent in native HIVTat monomer, as well as the general instability of peptides linkedtogether side by side as investigated, render this design unsuitable forin vivo use and clinical application owing to safety problems. DAGRS™use two inherent properties of Tat, one that locks a signal-transducingpeptide into an active conformation within the first 20 amino acids ofthe protein, making for the cloning cassette, and the well-described TARand RK-rich membrane translocation sequence (aa 38-60 of SF2 Tat). AnSH3 binding domain described here naturally encoded at the aminoterminus of HIV-1 Tat is removed from the cassette, and inhibitorypeptides for eg AKT or E2F are swapped into the cassette. As we havediscovered that this SH3-binding domain is a major source of toxicity,in this process DAGRS™ are rendered much safer than Tat. A CRD that sitsjust carboxyl to the cassette, and is a major source of Tat reactivity,is “humanized” to remove toxicity by replacement alternatively with ahuman C-rich spacer, or the CRD from non-pathogenic SIV Tat. Because thepresent invention appreciates that DAGRS™ affinity forsignal-transducing target is a property of its conformation, and thatDAGRS™ bioavailability depends upon membrane translocation, the DAGRS™described in the present invention preserves both of thesefunctionalities. These DAGRS™ constructs provide improved safety andbetter bioavailability for therapeutics in the treatment of cancer andother in vivo applications.

DESCRIPTION OF THE FIGURES

FIG. 1: Schematic of DAGRS™ constructs. Illustrated are the 3 geneticcomponents (“tiles”) of a DAGRS™ construct: 1) an NH₂ terminal cassetteencoding a signal transducing peptide (STP) designed to competitivelyinhibit an oncogene, a C-Rich spacer preferentially encoding 6 cysteineresidues, that facilitates proper folding of the DAGRS™, and a RK-richmembrane translocation sequence (MTS), SEQ ID 2) at the COOH terminusthat is preceded by sequences interacting with cyclin (TAR, SEQ ID1,underlined) or other proteins that translocate from the cytoplasm to thenucleus. The entire third cassette could also be replaced by fully humansequences, such as amino acids 277 to 291 from Atx-3 that mediatesinteraction with human VCP with nuclear translocation of the complex.(Boeddrich et al. EMBO J 25, 1547, 2006.) Although modelled after themolecular design of Tat, either 0 or 1 of three DAGRS™ domains containTat sequences, permitting the approximation that DAGRS™ hold an overallhomology to HIV Tat between 0-33%.

FIG. 2: Schematic showing the amino acid alignment of NH₂ terminalsequences of SIV Tats with homology to transcription factors.

PRALINE program was used to generate the alignments.

FIG. 3: Schematic showing the amino acid alignment of NH₂ terminalsequences of HIV and CPZ Tat containing a P-rich string with homology toSH3-binding domains.

FIG. 4: Monomer Tat transcriptional activation of the HIV ltr. A. Graphof induced luciferase activity. HeLa cells were transfected with an LTRluciferase reporter construct. 24 hours after transfection at a timewhen cells were approximately 75% confluent, cells were scrape-loadedwith 2 μg protein in the presence of 100 μM chloroquine as described byFrankel (Frankel AD1, Pabo CO. Cell. 1988 Dec. 23; 55(6):1189-93.) Afteran additional 24 hours luciferase activity was measured and is recordedas relative activity compared to sham-transfected HeLa cells. Theaverage of three independent experiments is shown. B. Western blot ofrecombinant Tat proteins synthesized in E. coli probed with polyclonalantibodies from mice receiving Tat immunotherapy, Monomeric Tat wasisolated directly from cracked cells ((Lane 1), or Tat proteins weresequentially re-annealed to trimer (Lanes 2,3) by incubation withpolyclonal antibody to aa 57-72 peptide (HIV1 SF2). Molecular weight inkilodaltons (MrKd). Trimeric Tat runs as an apparent 48 kd substrate andmonomer as 16 kd; wherever monomer is present at high concentrations,cysteine-linked dimers are also observed (Lane 1).

FIG. 5: Bioavailability of Tat after membrane translocation. Peripheralblood mononuclear cells (PBMCs, 10⁶ cells/ml) were activated withphytohemagglutinin (PHA, 10 μg/ml) for 48 hours. PBMCs were washed,re-suspended at 10⁷ cells/ml, and scrape-loaded with Tat geneticallylinked to Green Fluorescent Protein (Tat-GFP, 10 μM). PBMCs were dilutedback to 10⁶ cells/ml and cultured overnight (15 hours) in RPMI-10% FCSat which time they were harvested and assayed for green fluorescence.

FIG. 6: Compositions of Matter encoding oncogene-antagonizing peptides.

SEQ ID 6 PKPPSKKRSCDPSEYR

DAGRS™ peptide derived from chicken anemia virus VP3

SEQ ID 7 PPFKPQVTSETRYF

DAGRS™ peptide derived from the SH3-binding region of human AKT

SEQ ID 8 PPKPPQVTSETDTRYF

DAGRS™ peptide derived from AKT modified to be “right-handed” as VP3 andto contain a canonical PPxPP Src SH3-binding site. (Kay, Williamson, andSudol, FASEB J 14, 231, 2000).

SEQ ID 9 HHHRLSH

DAGRS™ encoding the E2F promoter binding peptide as described byBertino.

FIG. 7: Algorithm for Humanizing a viral peptide. in silicoidentification of the apoptotic determinant in CAV VP3 as a peptidehomologous to the SH3-binding domain of AKT. The alignment is anchoredby a P-rich SH3-binding region (Orange box) at its amino end, andanchored at its carboxyl end by an S/TxY (blue and green boxes) MAPkinase phosphorylation motif.

DETAILED DESCRIPTION OF THE INVENTION

DAGRS™ are targeted drugs aimed to control tumor growth, and prevent orresolve metastases, while avoiding many of the side effects associatedwith standard chemotherapy.

The present invention provides for improved oncogene-directed biologicswhich in its simplest realization locks a signal transducing peptideinto an NH₂ terminal cassette in a biologically active configurationprotected from degradation, and links this sequence to a carboxylKR-rich membrane translocation sequence (MTS, SEQ ID2). The construct isdesigned to facilitate bioavailability and stability of theoncogene-inhibitory peptide. The Tat-encoded membrane translocationsequence (SEQ ID2, “penetrin”) has been screened for safety in clinicaltrial (Voskens et al. Head Neck 34, 1734, 2012). Additionally, Tatcontains sequences critical for its entering the nuclear transcriptome(TAR) and to its binding cyclin (SEQ ID1, underlined) that are apreferred realization of this invention (SEQ ID 1), because they areproposed to maintain all functionalities while conserving correct domainspacing within Tat. It is not known whether TAR (aa 38-47 within Tat)contributes to Tat toxicity, so another realization of this inventionpreferred for safety replaces Tat TAR/MTS with a fully human sequencestudied to have similar functionalities as Tat. As an example, humanAtx-3 mediates the translocation of human VCP to the nucleus: thepeptide sequence responsible for these functionalities is illustrated(Atx-3 amino acids 277-291, SEQ ID 3). Noteworthy that like Tat SEQ ID3encodes a short stretch of amino acids preceding its KR-rich MTS. Aschematic of the DAGRS™ construct is illustrated in FIG. 1.

DAGRS™ use a molecular design evolved by the SIV/HIV Tat protein, butare humanized for safety. Overall, DAGRS™ composition of matter rangebetween 0-33% identity to Tat. This is critical because HIV Tat is atoxic substance which precludes its use in clinical applications.Following the molecular design of SIV Tat (FIG. 2) and HIV/CPZ Tat (FIG.3), DAGRS™ contain a signal transducing peptide (STP) at their NH₂terminus. The natural STP of the Tat design is removed, leaving behind acassette for inserting a peptide theoretically capable of inhibiting anyoncogene dependent on the sequence of the STP. An important embodimentof the present invention contemplates replacement of the Tat CRD ligandcombining site with a neutral human genetic spacer of relatively similarcomposition, and in particular containing 6 C residues to facilitateproper folding of the biologic. Two examples illustrating this inventionare the CRD (amino acids 41-60) from human β-defensin 4 (SEQ ID 4) orthe CRD from human wnt3 (SEQ ID 5), although any human CRD pairing 6 Cis a preferred realization of this invention. Without modification, theCRD region, which is very autoreactive, could cause major toxicity.Therefore the present invention has redesigned (“humanized”) the regionto provide for significantly less toxicity.

FIG. 2 shows an alignment of acidic transcription factor peptides (TFP)encoded at the NH₂ terminus of SIV. The sequences are stapled on oneside by the M initiator and move directly into a DE-rich (acidictranscription activator) segment. The SIV TFP are locked intoconfiguration by a conserved P at their COOH end just before the startof the CRD, as illustrated in FIG. 2. FIG. 3 demonstrates that HIV-1,HIV-2, and CPZ carry a different class of STP at their NH₂ terminus withcharacteristics of SH3-binding domains. In particular these three TatSTP initiate with a PxxP motif canonical for SH3-binding domains. As forSIV Tat, they are stapled at their COOH end by another P just prior tothe C-rich domain (FIG. 3).

FIG. 4 shows the well-known property of monomeric Tat to enter into andinfluence cellular transcription. The TAR region (SEQ ID 1, underlined)and the MTS are critical components of this functionality. Inparticular, HeLa cells loaded with 2 μg monomeric Tat proteintransactivated an ltr-luciferase construct 85 fold above sham-loadedHeLa cells, or HeLa loaded with 2 μg trimeric Tat, which is thebiologically active form for oncoimmunotherapeutics. Tat provides themeans to introduce the oncogene-inhibitory STP fragment at the site ofaction. FIG. 4 shows the activity of the monomer as a transcriptionalregulator.

The present invention further improves bioavailability by combining themembrane translocation sequences of Tat with the targeted killing effectof Apoptin, or any STP. The invention is not interfered with by patentfilings proposing to link Apoptin to Tat for improved bioavailability(US 20020176860, US 2008/0234466) because those inventors on theTat-Apoptin patents acknowledge that native Tat is too toxic to beadministered to humans (see Los et al., Apoptin, a tumor-selectivekiller, Biochem Biophys Acta. 1793, (2009) 1335-1342), and because theDAGRS™ sequences bear <33% identity to Tat. Translocation ofTat-GFP-monomer is shown in FIG. 5. Here, cells are incubated in 10micromolar Tat-GFP monomer and fluorescently imaged.

FIG. 6 details specific DAGRS™ compositions of matter to be embeddedinto the STD (SEQ ID 6-9). In particular, SEQ ID 6 proposes a DAGRS™with an unmodified 16 mer from the CAV VP3 (“Apoptin”) that contains theVP3 SH3-binding site and flanking carboxly amino acids. CAV VP3(“Apoptin”), has been shown to specifically interact with AKT. Apoptinis known to kill most cancers using an AKT pathway, but Apoptin is notcytotoxic for normal cells. Prior to the present invention, nothingsimilar has been described that could function as an in vivo oncolyticwithout associated toxicity. SEQ ID 7 identifies a 16 mer homologue tothe VP3 SH3-binding domain and carboxyl flanking sequences in human AKT,strongly supporting the proposal that VP3 functions as a competitiveinhibitor to AKT activation. Further, polio oncolytic virus, nowapproved for treating recurrent glioblastoma cancers, appears to work,at least in part, through an AKT mechanism. The present inventionproposes using the human AKT SH3-binding region (SEQ ID 7) as analternative to SEQ ID 6 under the proposition that its fully humansequence will be better tolerated and less immunogenic in the clinicalsetting. A third DAGRS™ derivative (SEQ ID 8) aligns the paired prolinesat the carboxyl terminus, as in VP3, but also retains the paired aminoprolines, and in so doing creates a DAGRS™ with a canonical SH3-bindingsite for the src oncogene. Interaction between Src and the SH3-bindingregion of AKT is requisite for AKT activation (Jiang and Qiu, Journal ofBiological Chemistry 278, 15789, 2003). SEQ ID8 describes aderivatization of AKT peptide that could give it higher affinity for Srcthan native human AKT. A further realization of the composition isextending 16 mer SEQ ID 6-8 with 4 or 5 COOH amino acids from theirrespective proteins (ie RVSEL for VP3), thereby engineering 20 mers withproposed carboxyl phosphorylation site anchors more distant from their Pstaples (FIG. 7), since the P staples are not present in either VP3 orAKT whole protein, and could be a source of steric hindrance.

Another embodiment of the present invention is a DAGRS™ with atranscription factor/protein activator region such as a peptide capableof binding to E2F promoter ((SEQ ID 9). This design bears analogy to SIVTat in encoding an acidic region, while the AKT design bears analogy toHIV Tat in encoding an SH3-binding domain As for SEQ ID 6-8, it could bebeneficial to distance E2F peptide (SEQ ID 9) by 4 or 5 amino acids fromthe COOH P staple.

FIG. 7 demonstrates the functional alignment between the viral VP3SH3-binding domain, and human AKT SH3-binding domain, and establishes analgorithm generally useful for the alignment of viral tiles with humantiles (a “humanization” algorithm). In particular other than thealignment of the proline-rich SH3-binding domains (boxed orange) thereis little or no homology (and none recognized by current standardprotein alignment programs) until the sequences reach a tightlyevolutionarily conserved S/TxY Mitogen Activated Protein kinasephosphorylation site (Mohanta et al Biological Procedures Online 17, 13,2015) at their COOH end, with an identical spacing of 9 amino acidsbetween P and S/T (boxed blue). Particularly insofar as AKT is known torequire Y phosphorylation (by Src) for activation, a reasonable model isthat in resting cells S/T is phosphorylated and sterically inhibiting Y(boxed green) phosphorylation carboxyl by 2 amino acids. Upon oncogenictransformation, which is known to induce phosphatases, S/T would becomedephosphorylated and Y susceptible to AKT oncogenic phosphorylation. Ineither case, the VP3 cassette (SEQ ID 6) could function as a competitiveinhibitor to AKT activation, as would be also expected for the Yphosphorylation sites of SEQ ID 7 and 8.

This is the first time that a functional viral domain has been matchedup (“humanized”) to a human protein fragment, and in so doing describesa key humanization invention. The example of FIG. 7 can be generalizedto an algorithm that identifies conserved functionalities among proteins(AKT and VP3 both influence G2 phase cell cycle transition and interactwith identical proteins), aligns peptides from the two proteins throughamino and carboxyl anchors with matched functional domains, eg anSH3-binding domain and a MAP kinase phosphorylation domain, andprioritizes equivalent spacing between the two functional domains, whileat the same time totally ignoring the primary amino acid sequence of thespacer. The rationale supporting the algorithm is that correct aminoacid spacing conserves functional interactions in three dimensions. Thealgorithm totally ignores the composition of amino acids interveningbetween the anchors, because the evolutionary distance between thespecies originally hosting the virus and humans is proposed to be toodistant to conserve primary amino acid sequence of these non-essentialresidues. This is consistent with the vectors that transmit Zika virusand Ebola virus to humans, being respectively mosquitoes and bats. Manyviral activities could be rendered safe and therapeutic via thisalgorithm converting viral components to human protein components.

Although the present invention has been described with reference tospecific embodiments, workers skilled in the art will recognize thatmany variations may be made therefrom and it is to be understood andappreciated that the disclosures in accordance with the invention showonly some preferred embodiments and advantages of the invention withoutdeparting from the broader scope and spirit of the invention. It is tobe understood and appreciated that these discoveries in accordance withthis invention are only those which are illustrated of the manyadditional potential applications that may be envisioned by one ofordinary skill in the art, and thus are not in any way intended to belimiting of the invention. Accordingly, other objects and advantages ofthe invention will be apparent to those skilled in the art from thedetailed description together with the claims.

SEQ ID No Peptide sequence Source 1 FTRKGLGISYGRKKRRQRRR HIV 2 RKKRRQRRRHUMAN 3 TSEELRKRREAYFEK HUMAN 4 CLTKGGVCWGPCTGGFRQIGTCGLPRVRCC HUMAN 5CRCVFHWCCYVSCQEC HUMAN 6 PKPPSKKRSCDPSEYR CHICKEN  ANEMIA VIRUS 7PPFKPQVTSETDTRYF HUMAN 8 PPKPPQVTSETDTRYF HUMAN 9 HHHRLSH HUMAN

What is claimed is:
 1. A method for treating cancer in a patientcomprising administering a therapeutically effective amount of a DAGRS™construct having at its amino cassette a signal transducing peptide(STP) able to inhibit activity of a specific oncogene, followed by aC-rich spacer, and further followed by a sequence enabling nuclearprotein interactions and membrane translocations wherein the saidadministration causes a cessation of growth or regression of said cancerin said patient.
 2. The DAGRS™ of claim 1 with its amino cassettecomprising any peptide inhibiting the AKT or E2F oncogenes.
 3. The AKTpeptide of claim 2 (SEQ ID 8) derivatized for enhanced affinity to theSrc oncogene.
 4. The peptide of claim 1 inserted into a DAGRS™ cassettefunctioning to inhibit any oncogene.
 5. The DAGRS™ construct of claim 1consisting of an inhibitory peptide for AKT, E2F, CRK, or any oncogene,followed by a cysteine-rich spacer, followed by a nucleartranslocation/interaction peptide.
 6. The DAGRS™ of claim 1 with a 6Cysteine spacer.
 7. The DAGRS™ of claim 1 with a 6 Cysteine spacerderived from the βdefensin or wnt family of proteins.
 8. The DAGRS™ ofclaim 1 with a spacer approximating 15 amino acids of any amino acidcomposition.
 9. The DAGRS™ of claim 1 where the sequence enablingnuclear interactions and membrane translocations is the TAR/MTSsequences (aa 38-57, SEQ ID 1) of HIV or SIV.
 10. The DAGRS™ of claim 1where the sequence enabling nuclear interactions and membranetranslocations is the sequence (aa 271-291, SEQ ID 3) of Atx-3.
 11. TheDAGRS™ of claim 1 with a penetrin sequence (SEQ ID 2) at its carboxylterminus.
 12. A method for removing toxicity from viral products(“humanizing”) by identifying human peptides with equivalent functionalactivity using an algorithm.
 13. The algorithm of claim 12 that matchesup functionally equivalent peptides between viral and human amino acidsequences.
 14. The algorithm of claim 13 that matches functionallyequivalent peptides by anchoring the alignment with identical motifs atits amino and carboxyl ends, and identifying identical spacing of themotifs, without regard to the primary sequence of the amino acidsintervening between the two anchors.
 15. The algorithm of claim 14wherein Apoptin and AKT are aligned by an amino anchor of an SH3-bindingmotif compatible with Src oncogene interaction, and a carboxyl anchor ofa MAP kinase phosphorylation site.
 16. The algorithm of claim 14 furtherhaving a computer program to humanize viral activities.
 17. The use ofartificial intelligence to enhance the algorithm of claim
 14. 18. Thealgorithm of claim 14 used to discover oncogene-inhibitory peptides,oncolytic drugs or oncoimmunologic drugs.
 19. The use of the algorithmof claim 14 to develop drugs from humanized viral sequences for treatingAlzheimer's disease, cardiac arrhythmias, fever, rheumatoid arthritis orother autoimmune diseases.
 20. The use of the algorithm of claim 14 todevelop drugs for treating human diseases.
 21. The use of the algorithmof claim 14 used to develop compounds that alter the normal course ofaging.